Co-administration of rosiglitazone and eicosapentaenoic acid or a derivative thereof

ABSTRACT

In various embodiments, the present invention provides methods of treating and/or preventing cardiovascular-related disease and, in particular, a method of reducing triglycerides in a subject on rosiglitazone therapy, the method comprising administering to a subject in need thereof a pharmaceutical composition comprising eicosapentaenoic acid or a derivative thereof.

PRIORITY CLAIM

This application claims priority to U.S. Provisional patent applicationSer. No. 61/832,057, filed Jun. 6, 2013, and U.S. Provisional patentapplication Ser. No. 61/875,779, filed Sep. 10, 2013, the entirecontents of each of which are incorporated herein by reference andrelied upon.

BACKGROUND

Cardiovascular disease is one of the leading causes of death in theUnited States and most European countries. It is estimated that over 70million people in the United States alone suffer from a cardiovasculardisease or disorder including but not limited to high blood pressure,coronary heart disease, dyslipidemia, congestive heart failure andstroke. A need exists for improved treatments for cardiovasculardiseases and disorders.

SUMMARY

In various embodiments, the present invention provides methods oftreating and/or preventing cardiovascular-related diseases and, inparticular, a method of treating hypercholesterolemia comprisingadministering to a subject in need thereof a pharmaceutical compositioncomprising eicosapentaenoic acid or a derivative thereof. In oneembodiment, the composition is co-administered with rosiglitazone. Inone embodiment, the composition contains not more than 10%, by weight,of all fatty acids (and/or derivatives thereof) present, docosahexaenoicacid or derivative thereof, substantially no docosahexaenoic acid orderivative thereof, or no docosahexaenoic acid or derivative thereof. Inanother embodiment, eicosapentaenoic acid ethyl ester comprises at least96%, by weight, of all fatty acids (and/or derivatives thereof) presentin the composition; the composition contains not more than 4%, byweight, of total fatty acids (and/or derivatives thereof) other thaneicosapentaenoic acid ethyl ester; and/or the composition contains about0.1% to about 0.6% of at least one fatty acid other thaneicosapentaenoic acid ethyl ester and docosahexaenoic acid (orderivative thereof).

In one embodiment, the present disclosure provides a method of reducingtriglycerides in a subject on rosiglitazone therapy, the methodcomprising administering to the subject a pharmaceutical compositioncomprising at least about 80%, by weight of all fatty acids (and/orderivatives thereof) present, ethyl eicosapentaenoate.

In one embodiment, the present disclosure provides a method of reducingtriglycerides in a subject on rosiglitazone therapy, the methodcomprising administering to the subject about 4 g per day of ethyleicosapentaenoate.

In one embodiment, the present disclosure provides a method of reducingtriglycerides in a subject in need thereof, the method comprising,co-administering rosiglitazone and about 2 g or about 4 g per day ofethyl eicosapentaenoate, wherein said co-administration provides asteady state plasma C_(max) and/or a steady state plasma AUC_(0-inf) ofrosiglitazone of about 70% to about 135% of a mean steady state plasmaC_(max) and/or a mean steady state plasma AUC_(0-inf) of rosiglitazonein subjects receiving said rosiglitazone daily without the ethyleicosapentaenoate.

In one embodiment, the present disclosure provides a method of reducinga risk of a cardiovascular event in a subject on rosiglitazone therapy,the method comprising administering to the subject a pharmaceuticalcomposition comprising at least about 80%, by weight of all fatty acids(and/or derivatives thereof) present, ethyl eicosapentaenoate.

In one embodiment, the present disclosure provides a method of reducinga risk of a cardiovascular event in a subject on rosiglitazone therapy,the method comprising in a subject on rosiglitazone therapy, the methodcomprising administering to the subject about 4 g per day of ethyleicosapentaenoate.

In one embodiment, a pharmaceutical composition useful in accordancewith the invention comprises, consists of or consists essentially of atleast 95%, by weight of all fatty acids (and/or derivatives thereof)present, ethyl eicosapentaenoate (EPA-E), about 0.2% to about 0.5%, byweight of all fatty acids (and/or derivatives thereof) present, ethyloctadecatetraenoate (ODTA-E), about 0.05% to about 0.25%, by weight ofall fatty acids (and/or derivatives thereof) present, ethylnonadecapentaenoate (NDPA-E), about 0.2% to about 0.45%, by weight ofall fatty acids (and/or derivatives thereof) present, ethyl arachidonate(AA-E), about 0.3% to about 0.5%, by weight of all fatty acids (and/orderivatives thereof) present, ethyl eicosatetraenoate (ETA-E), and about0.05% to about 0.32%, by weight of all fatty acids (and/or derivativesthereof) present, ethyl heneicosapentaenoate (HPA-E). In anotherembodiment, the composition is present in a capsule shell. In anotherembodiment, the composition contains substantially no or no amount ofdocosahexaenoic acid (DHA) or derivative thereof such as ethyl-DHA(DHA-E).

In another embodiment, the invention provides a method of treatingmoderate to severe hypertriglyceridemia comprising administering acomposition as described herein to a subject in need thereof one toabout four times per day.

In some embodiments, the present invention comprises co-administered ofethyl eicosapentaenoate with rosiglitazone. In some embodiments,co-administration of 2 g or 4 g of ethyl eicosapentaenoate androsiglitazone provides a mean steady state plasma C_(max), a mean steadystate plasma AUC_(0-inf), and/or a mean steady state plasma T_(max) ofrosiglitazone of about 70% to about 135% of a mean steady state plasmaC_(max), a mean steady state plasma AUC_(0-inf), and/or a mean steadystate plasma T_(max) of rosiglitazone when rosiglitazone is administeredto subjects without the ethyl eicosapentaenoate.

In some embodiments, a composition of the present invention comprises atleast about 80%, by weight of all fatty acids (and/or derivativesthereof) present, ethyl eicosapentaenoate, wherein the composition doesnot significantly alter a blood plasma C_(max), a blood plasmaAUC_(0-inf), and/or a blood plasma T_(max) of rosiglitazone.

In some embodiments, a method of reducing triglycerides in a subject onrosiglitazone therapy according to the present invention comprisesadministering to the subject a composition comprising at least about80%, by weight of all fatty acids (and/or derivatives thereof) present,ethyl eicosapentaenoate.

These and other embodiments of the present invention will be disclosedin further detail herein below.

BRIEF DESCRIPTION OF THE FIGURE

FIG. 1 shows mean plasma concentrations of rosigilitazone over time whenadministered with 4 grams/day of ethyl eicosapentaenoate (squares) andwithout ethyl eicosapentaenoate (diamonds).

DETAILED DESCRIPTION

While the present invention is capable of being embodied in variousforms, the description below of several embodiments is made with theunderstanding that the present disclosure is to be considered as anexemplification of the invention, and is not intended to limit theinvention to the specific embodiments illustrated. Headings are providedfor convenience only and are not to be construed to limit the inventionin any manner. Embodiments illustrated under any heading may be combinedwith embodiments illustrated under any other heading.

The use of numerical values in the various quantitative values specifiedin this application, unless expressly indicated otherwise, are stated asapproximations as though the minimum and maximum values within thestated ranges were both preceded by the word “about.” Also, thedisclosure of ranges is intended as a continuous range including everyvalue between the minimum and maximum values recited as well as anyranges that can be formed by such values. Also disclosed herein are anyand all ratios (and ranges of any such ratios) that can be formed bydividing a disclosed numeric value into any other disclosed numericvalue. Accordingly, the skilled person will appreciate that many suchratios, ranges, and ranges of ratios can be unambiguously derived fromthe numerical values presented herein and in all instances such ratios,ranges, and ranges of ratios represent various embodiments of thepresent invention.

In one embodiment, the invention provides a method for treatment and/orprevention of a cardiovascular-related disease. The term“cardiovascular-related disease” herein refers to any disease ordisorder of the heart or blood vessels (i.e. arteries and veins) or anysymptom thereof. Non-limiting examples of cardiovascular-related diseaseand disorders include hypertriglyceridemia, hypercholesterolemia, mixeddyslipidemia, coronary heart disease, vascular disease, stroke,atherosclerosis, arrhythmia, hypertension, myocardial infarction, andother cardiovascular events.

The term “treatment” in relation a given disease or disorder, includes,but is not limited to, inhibiting the disease or disorder, for example,arresting the development of the disease or disorder; relieving thedisease or disorder, for example, causing regression of the disease ordisorder; or relieving a condition caused by or resulting from thedisease or disorder, for example, relieving, preventing or treatingsymptoms of the disease or disorder. The term “prevention” in relationto a given disease or disorder means: preventing the onset of diseasedevelopment if none had occurred, preventing the disease or disorderfrom occurring in a subject that may be predisposed to the disorder ordisease but has not yet been diagnosed as having the disorder ordisease, and/or preventing further disease/disorder development ifalready present.

Rosiglitazone is a thiazolidinedione insulin sensitizing drug which wasfound to increase the risk of heart attacks, despite its efficacy as adiabetic therapy. It is a cytochrome P450 2C8 (“CYP2C8”) substrate. Thedaily dose of rosiglitazone is typically 4 mg or 8 mg, taken in singleor in divided doses. It has an empirical chemical formula ofC₁₈H₁₉N₃O₃S, a molecular weight of 357.43 g/mol, and the structure shownbelow:

As used herein, the term “rosiglitazone” refers to the compound shownand described above, as well as any therapeutic derivatives or formsthereof, such as an enriched or purified enantiomer thereof.

In one embodiment, the present invention provides a method of bloodlipid therapy comprising administering to a subject or subject group inneed thereof a pharmaceutical composition as described herein. Inanother embodiment, the subject or subject group has one or more of:hypercholesterolemia, familial hypercholesterolemia, high LDL-C serumlevels, high total cholesterol levels, and/or low HDL-C serum levels.

In another embodiment, the subject or subject group being treated has abaseline triglyceride level (or median baseline triglyceride level inthe case of a subject group), fed or fasting, of at least about 300mg/dl, at least about 400 mg/dl, at least about 500 mg/dl, at leastabout 600 mg/dl, at least about 700 mg/dl, at least about 800 mg/dl, atleast about 900 mg/dl, at least about 1000 mg/dl, at least about 1100mg/dl, at least about 1200 mg/dl, at least about 1300 mg/dl, at leastabout 1400 mg/dl, or at least about 1500 mg/dl, for example about 400mg/dl to about 2500 mg/dl, about 450 mg/dl to about 2000 mg/dl or about500 mg/dl to about 1500 mg/dl.

In one embodiment, the subject or subject group being treated inaccordance with methods of the invention has previously been treatedwith Lovaza® and has experienced an increase in, or no decrease in,LDL-C levels and/or non-HDL-C levels. In one such embodiment, Lovaza®therapy is discontinued and replaced by a method of the presentinvention.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineabsolute plasma level of free EPA (or mean thereof in the case of asubject group) not greater than about 0.70 nmol/ml, not greater thanabout 0.65 nmol/ml, not greater than about 0.60 nmol/ml, not greaterthan about 0.55 nmol/ml, not greater than about 0.50 nmol/ml, notgreater than about 0.45 nmol/ml, or not greater than about 0.40 nmol/ml.In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a baseline fastingplasma level (or mean thereof) of free EPA, expressed as a percentage oftotal free fatty acid, of not more than about 3%, not more than about2.5%, not more than about 2%, not more than about 1.5%, not more thanabout 1%, not more than about 0.75%, not more than about 0.5%, not morethan about 0.25%, not more than about 0.2% or not more than about 0.15%.In one such embodiment, free plasma EPA and/or total fatty acid levelsare determined prior to initiating therapy.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineabsolute plasma level of total fatty acid (or mean thereof) not greaterthan about 250 nmol/ml, not greater than about 200 nmol/ml, not greaterthan about 150 nmol/ml, not greater than about 100 nmol/ml, or notgreater than about 50 nmol/ml.

In another embodiment, the subject or subject group being treated inaccordance with methods of the invention exhibits a fasting baselineplasma, serum or red blood cell membrane EPA level not greater thanabout 70 μg/ml, not greater than about 60 μg/ml, not greater than about50 μg/ml, not greater than about 40 μg/ml, not greater than about 30μg/ml, or not greater than about 25 μg/ml.

In another embodiment, methods of the present invention comprise a stepof measuring the subject's (or subject group's mean) baseline lipidprofile prior to initiating therapy. In another embodiment, methods ofthe invention comprise the step of identifying a subject or subjectgroup having one or more of the following: baseline non-HDL-C value ofabout 200 mg/dl to about 400 mg/dl, for example at least about 210mg/dl, at least about 220 mg/dl, at least about 230 mg/dl, at leastabout 240 mg/dl, at least about 250 mg/dl, at least about 260 mg/dl, atleast about 270 mg/dl, at least about 280 mg/dl, at least about 290mg/dl, or at least about 300 mg/dl; baseline total cholesterol value ofabout 250 mg/dl to about 400 mg/dl, for example at least about 260mg/dl, at least about 270 mg/dl, at least about 280 mg/dl or at leastabout 290 mg/dl; baseline vLDL-C value of about 140 mg/dl to about 200mg/dl, for example at least about 150 mg/dl, at least about 160 mg/dl,at least about 170 mg/dl, at least about 180 mg/dl or at least about 190mg/dl; baseline HDL-C value of about 10 to about 60 mg/dl, for examplenot more than about 40 mg/dl, not more than about 35 mg/dl, not morethan about 30 mg/dl, not more than about 25 mg/dl, not more than about20 mg/dl, or not more than about 15 mg/dl; and/or baseline LDL-C valueof about 50 to about 300 mg/dl, for example not less than about 100mg/dl, not less than about 90 mg/dl, not less than about 80 mg/dl, notless than about 70 mg/dl, not less than about 60 mg/dl or not less thanabout 50 mg/dl.

In a related embodiment, upon treatment in accordance with the presentinvention, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits one or more of the following outcomes:

(a) reduced triglyceride levels compared to baseline or control;

(b) reduced Apo B levels compared to baseline or control;

(c) increased HDL-C levels compared to baseline or control;

(d) no increase in LDL-C levels compared to baseline or control;

(e) a reduction in LDL-C levels compared to baseline or control;

(f) a reduction in non-HDL-C levels compared to baseline or control;

(g) a reduction in vLDL levels compared to baseline or control;

(h) an increase in apo A-I levels compared to baseline or control;

(i) an increase in apo A-Papo B ratio compared to baseline or control;

(j) a reduction in lipoprotein A levels compared to baseline or control;

(k) a reduction in LDL particle number compared to baseline or control;

(l) an increase in LDL size compared to baseline or control;

(m) a reduction in remnant-like particle cholesterol compared tobaseline or control;

(n) a reduction in oxidized LDL compared to baseline or control;

(o) no change or a reduction in fasting plasma glucose (FPG) compared tobaseline or control;

(p) a reduction in hemoglobin A_(lc) (HbA_(lc)) compared to baseline orcontrol;

(q) a reduction in homeostasis model insulin resistance compared tobaseline or control;

(r) a reduction in lipoprotein associated phospholipase A2 compared tobaseline or control;

(s) a reduction in intracellular adhesion molecule-1 compared tobaseline or control;

(t) a reduction in interleukin-6 compared to baseline or control;

(u) a reduction in plasminogen activator inhibitor-1 compared tobaseline or control;

(v) a reduction in high sensitivity C-reactive protein (hsCRP) comparedto baseline or control;

(w) an increase in serum or plasma EPA compared to baseline or control;

(x) an increase in red blood cell (RBC) membrane EPA compared tobaseline or control;

(y) a reduction or increase in one or more of serum phospholipid and/orred blood cell content of docosahexaenoic acid (DHA), docosapentaenoicacid (DPA), arachidonic acid (AA), palmitic acid (PA), staeridonic acid(SA) or oleic acid (OA) compared to baseline or control;

(z) a reduction in or prevention of membrane cholesterol domainformation compared to baseline or control; and/or

(aa) a reduction in or prevention of oxidative modification of membranepolyunsaturated fatty acids compared to baseline or control.

In one embodiment, upon administering a composition of the invention toa subject, the subject exhibits a decrease in triglyceride levels, anincrease in the concentrations of EPA and DPA (n−3) in red blood cells,and an increase of the ratio of EPA:arachidonic acid in red blood cells.In a related embodiment the subject exhibits substantially no or noincrease in RBC DHA.

In one embodiment, methods of the present invention comprise measuringbaseline levels of one or more markers set forth in (a)-(aa) above priorto dosing the subject or subject group. In another embodiment, themethods comprise administering a composition as disclosed herein to thesubject after baseline levels of one or more markers set forth in(a)-(aa) are determined, and subsequently taking an additionalmeasurement of said one or more markers.

In another embodiment, upon treatment with a composition of the presentinvention, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits any 2 or more of, any 3 or more of,any 4 or more of, any 5 or more of, any 6 or more of, any 7 or more of,any 8 or more of, any 9 or more of, any 10 or more of, any 11 or moreof, any 12 or more of, any 13 or more of, any 14 or more of, any 15 ormore of, any 16 or more of, any 17 or more of, any 18 or more of, any 19or more of, any 20 or more of, any 21 or more of, any 22 or more of, any23 or more, any 24 or more, any 25 or more, any 26 or more, or all 27 ofoutcomes (a)-(aa) described immediately above.

In another embodiment, upon treatment with a composition of the presentinvention, the subject or subject group exhibits one or more of thefollowing outcomes:

(a) a reduction in triglyceride level of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) as compared to baseline;

(b) a less than 30% increase, less than 20% increase, less than 10%increase, less than 5% increase or no increase in non-HDL-C levels or areduction in non-HDL-C levels of at least about 1%, at least about 3%,at least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55% or at least about 75% (actual % change or median % change) ascompared to baseline;

(c) substantially no change in HDL-C levels, no change in HDL-C levels,or an increase in HDL-C levels of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, at least about 55% or at least about 75% (actual %change or median % change) as compared to baseline;

(d) a less than 60% increase, a less than 50% increase, a less than 40%increase, a less than 30% increase, less than 20% increase, less than10% increase, less than 5% increase or no increase in LDL-C levels or areduction in LDL-C levels of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, at least about 55%, at least about 55% or at leastabout 75% (actual % change or median % change) as compared to baseline;

(e) a decrease in Apo B levels of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, at least about 55% or at least about 75% (actual %change or median % change) as compared to baseline;

(f) a reduction in vLDL levels of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, or at least about 100% (actual % change or median %change) compared to baseline;

(g) an increase in apo A-I levels of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline;

(h) an increase in apo A-I/apo B ratio of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline;

(i) a reduction in lipoprotein (a) levels of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline;

(j) a reduction in mean LDL particle number of at least about 5%, atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline;

(k) an increase in mean LDL particle size of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, or at least about 100% (actual % changeor median % change) compared to baseline;

(l) a reduction in remnant-like particle cholesterol of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline;

(m) a reduction in oxidized LDL of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline;

(n) substantially no change, no significant change, or a reduction (e.g.in the case of a diabetic subject) in fasting plasma glucose (FPG) of atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, or at leastabout 100% (actual % change or median % change) compared to baseline;

(o) substantially no change, no significant change or a reduction inhemoglobin A_(lc) (HbA_(lc)) of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,or at least about 50% (actual % change or median % change) compared tobaseline;

(p) a reduction in homeostasis model index insulin resistance of atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, or at leastabout 100% (actual % change or median % change) compared to baseline;

(q) a reduction in lipoprotein associated phospholipase A2 of at leastabout 5%, at least about 10%, at least about 15%, at least about 20%, atleast about 25%, at least about 30%, at least about 35%, at least about40%, at least about 45%, at least about 50%, or at least about 100%(actual % change or median % change) compared to baseline;

(r) a reduction in intracellular adhesion molecule-1 of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline;

(s) a reduction in interleukin-6 of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, or at least about 100% (actual % change ormedian % change) compared to baseline;

(t) a reduction in plasminogen activator inhibitor-1 of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, or at least about 100% (actual %change or median % change) compared to baseline;

(u) a reduction in high sensitivity C-reactive protein (hsCRP) of atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, or at leastabout 100% (actual % change or median % change) compared to baseline;

(v) an increase in serum, plasma and/or RBC EPA of at least about 5%, atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, at least about 50%, at least about 100%, at least about200% or at least about 400% (actual % change or median % change)compared to baseline;

(w) an increase in serum phospholipid and/or red blood cell membrane EPAof at least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, r at least about 50%, at leastabout 100%, at least about 200%, or at least about 400% (actual % changeor median % change) compared to baseline;

(x) a reduction or increase in one or more of serum phospholipid and/orred blood cell DHA, DPA, AA, PA and/or OA of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) compared to baseline;

(y) a reduction in total cholesterol of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55% or at least about 75%(actual % change or median % change) compared to baseline;

(z) a reduction in membrane cholesterol domain formation of at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55%, at least about 60%,at least about 65%, at least about 70%, at least about 75%, at leastabout 80%, at least about 85%, at least about 90%, at least about 95%,at least about 98%, at least about 99%, or about 100% (actual % changeor median % change) compared to baseline or control; and/or

(aa) a reduction in oxidative modification of membrane polyunsaturatedfatty acids of at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, at least about55%, at least about 60%, at least about 65%, at least about 70%, atleast about 75%, at least about 80%, at least about 85%, at least about90%, at least about 95%, at least about 98%, at least about 99%, orabout 100% (actual % change or median % change) compared to baseline orcontrol.

In one embodiment, methods of the present invention comprise measuringbaseline levels of one or more markers set forth in (a)-(aa) prior todosing the subject or subject group. In another embodiment, the methodscomprise administering a composition as disclosed herein to the subjectafter baseline levels of one or more markers set forth in (a)-(aa) aredetermined, and subsequently taking a second measurement of the one ormore markers as measured at baseline for comparison thereto.

In another embodiment, upon treatment with a composition of the presentinvention, for example over a period of about 1 to about 200 weeks,about 1 to about 100 weeks, about 1 to about 80 weeks, about 1 to about50 weeks, about 1 to about 40 weeks, about 1 to about 20 weeks, about 1to about 15 weeks, about 1 to about 12 weeks, about 1 to about 10 weeks,about 1 to about 5 weeks, about 1 to about 2 weeks or about 1 week, thesubject or subject group exhibits any 2 or more of, any 3 or more of,any 4 or more of, any 5 or more of, any 6 or more of, any 7 or more of,any 8 or more of, any 9 or more of, any 10 or more of, any 11 or moreof, any 12 or more of, any 13 or more of, any 14 or more of, any 15 ormore of, any 16 or more of, any 17 or more of, any 18 or more of, any 19or more of, any 20 or more of, any 21 or more of, any 22 or more of, any23 or more of, any 24 or more of, any 25 or more of, any 26 or more of,or all 27 of outcomes (a)-(aa) described immediately above.

Parameters (a)-(y) can be measured in accordance with any clinicallyacceptable methodology. For example, triglycerides, total cholesterol,HDL-C and fasting blood sugar can be sample from serum and analyzedusing standard photometry techniques. VLDL-TG, LDL-C and VLDL-C can becalculated or determined using serum lipoprotein fractionation bypreparative ultracentrifugation and subsequent quantitative analysis byrefractometry or by analytic ultracentrifugal methodology. Apo Al, Apo Band hsCRP can be determined from serum using standard nephelometrytechniques. Lipoprotein (a) can be determined from serum using standardturbidimetric immunoassay techniques. LDL particle number and particlesize can be determined using nuclear magnetic resonance (NMR)spectrometry. Remnants lipoproteins and LDL-phospholipase A2 can bedetermined from EDTA plasma or serum and serum, respectively, usingenzymatic immunoseparation techniques. Oxidized LDL, intercellularadhesion molecule-1 and interleukin-6 levels can be determined fromserum using standard enzyme immunoassay techniques. These techniques aredescribed in detail in standard textbooks, for example TietzFundamentals of Clinical Chemistry, 6^(th) Ed. (Burtis, Ashwood andBorter Eds.), WB Saunders Company. Parameters (z) and (aa) can bemeasured in accordance with any clinically acceptable methodology or canbe estimated by any suitable in vitro experiment, for example, onesimilar to that described in Example 3.

In one embodiment, subjects fast for up to 12 hours prior to bloodsample collection, for example about 10 hours.

In another embodiment, the present invention provides a method oftreating or preventing primary hypercholesterolemia and/or mixeddyslipidemia (Fredrickson Types IIa and IIb) in a patient in needthereof, comprising administering to the patient one or morecompositions as disclosed herein. In a related embodiment, the presentinvention provides a method of reducing triglyceride levels in a subjector subjects when treatment with a statin or niacin extended-releasemonotherapy is considered inadequate (Frederickson type IVhyperlipidemia).

In another embodiment, the present invention provides a method oftreating or preventing risk of recurrent nonfatal myocardial infarctionin a patient with a history of myocardial infarction, comprisingadministering to the patient one or more compositions as disclosedherein.

In another embodiment, the present invention provides a method ofslowing progression of or promoting regression of atheroscleroticdisease in a patient in need thereof, comprising administering to asubject in need thereof one or more compositions as disclosed herein.

In another embodiment, the present invention provides a method oftreating or preventing very high serum triglyceride levels (e.g. TypesIV and V hyperlipidemia) in a patient in need thereof, comprisingadministering to the patient one or more compositions as disclosedherein.

In another embodiment, the present invention provides a method oftreating subjects having very high serum triglyceride levels (e.g.greater than 1000 mg/dl or greater than 2000 mg/dl) and that are at riskof developing pancreatitis, comprising administering to the patient oneor more compositions as disclosed herein.

In one embodiment, a composition of the invention is administered to asubject in an amount sufficient to provide a daily dose ofeicosapentaenoic acid of about 1 mg to about 10,000 mg, 25 about 5000mg, about 50 to about 3000 mg, about 75 mg to about 2500 mg, or about100 mg to about 1000 mg, for example about 75 mg, about 100 mg, about125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, about500 mg, about 525 mg, about 550 mg, about 575 mg, about 600 mg, about625 mg, about 650 mg, about 675 mg, about 700 mg, about 725 mg, about750 mg, about 775 mg, about 800 mg, about 825 mg, about 850 mg, about875 mg, about 900 mg, about 925 mg, about 950 mg, about 975 mg, about1000 mg, about 1025 mg, about 1050 mg, about 1075 mg, about 1100 mg,about 1025 mg, about 1050 mg, about 1075 mg, about 1200 mg, about 1225mg, about 1250 mg, about 1275 mg, about 1300 mg, about 1325 mg, about1350 mg, about 1375 mg, about 1400 mg, about 1425 mg, about 1450 mg,about 1475 mg, about 1500 mg, about 1525 mg, about 1550 mg, about 1575mg, about 1600 mg, about 1625 mg, about 1650 mg, about 1675 mg, about1700 mg, about 1725 mg, about 1750 mg, about 1775 mg, about 1800 mg,about 1825 mg, about 1850 mg, about 1875 mg, about 1900 mg, about 1925mg, about 1950 mg, about 1975 mg, about 2000 mg, about 2025 mg, about2050 mg, about 2075 mg, about 2100 mg, about 2125 mg, about 2150 mg,about 2175 mg, about 2200 mg, about 2225 mg, about 2250 mg, about 2275mg, about 2300 mg, about 2325 mg, about 2350 mg, about 2375 mg, about2400 mg, about 2425 mg, about 2450 mg, about 2475 mg, about 2500 mg,about 2525 mg, about 2550 mg, about 2575 mg, about 2600 mg, about 2625mg, about 2650 mg, about 2675 mg, about 2700 mg, about 2725 mg, about2750 mg, about 2775 mg, about 2800 mg, about 2825 mg, about 2850 mg,about 2875 mg, about 2900 mg, about 2925 mg, about 2950 mg, about 2975mg, about 3000 mg, about 3025 mg, about 3050 mg, about 3075 mg, about3100 mg, about 3125 mg, about 3150 mg, about 3175 mg, about 3200 mg,about 3225 mg, about 3250 mg, about 3275 mg, about 3300 mg, about 3325mg, about 3350 mg, about 3375 mg, about 3400 mg, about 3425 mg, about3450 mg, about 3475 mg, about 3500 mg, about 3525 mg, about 3550 mg,about 3575 mg, about 3600 mg, about 3625 mg, about 3650 mg, about 3675mg, about 3700 mg, about 3725 mg, about 3750 mg, about 3775 mg, about3800 mg, about 3825 mg, about 3850 mg, about 3875 mg, about 3900 mg,about 3925 mg, about 3950 mg, about 3975 mg, about 4000 mg, about 4025mg, about 4050 mg, about 4075 mg, about 4100 mg, about 4125 mg, about4150 mg, about 4175 mg, about 4200 mg, about 4225 mg, about 4250 mg,about 4275 mg, about 4300 mg, about 4325 mg, about 4350 mg, about 4375mg, about 4400 mg, about 4425 mg, about 4450 mg, about 4475 mg, about4500 mg, about 4525 mg, about 4550 mg, about 4575 mg, about 4600 mg,about 4625 mg, about 4650 mg, about 4675 mg, about 4700 mg, about 4725mg, about 4750 mg, about 4775 mg, about 4800 mg, about 4825 mg, about4850 mg, about 4875 mg, about 4900 mg, about 4925 mg, about 4950 mg,about 4975 mg, about 5000 mg, about 5025 mg, about 5050 mg, about 5075mg, about 5100 mg, about 5125 mg, about 5150 mg, about 5175 mg, about5200 mg, about 5225 mg, about 5250 mg, about 5275 mg, about 5300 mg,about 5325 mg, about 5350 mg, about 5375 mg, about 5400 mg, about 5425mg, about 5450 mg, about 5475 mg, about 5500 mg, about 5525 mg, about5550 mg, about 5575 mg, about 5600 mg, about 5625 mg, about 5650 mg,about 5675 mg, about 5700 mg, about 5725 mg, about 5750 mg, about 5775mg, about 5800 mg, about 5825 mg, about 5850 mg, about 5875 mg, about5900 mg, about 5925 mg, about 5950 mg, about 5975 mg, about 6000 mg,about 6025 mg, about 6050 mg, about 6075 mg, about 6100 mg, about 6125mg, about 6150 mg, about 6175 mg, about 6200 mg, about 6225 mg, about6250 mg, about 6275 mg, about 6300 mg, about 6325 mg, about 6350 mg,about 6375 mg, about 6400 mg, about 6425 mg, about 6450 mg, about 6475mg, about 6500 mg, about 6525 mg, about 6550 mg, about 6575 mg, about6600 mg, about 6625 mg, about 6650 mg, about 6675 mg, about 6700 mg,about 6725 mg, about 6750 mg, about 6775 mg, about 6800 mg, about 6825mg, about 6850 mg, about 6875 mg, about 6900 mg, about 6925 mg, about6950 mg, about 6975 mg, about 7000 mg, about 7025 mg, about 7050 mg,about 7075 mg, about 7100 mg, about 7125 mg, about 7150 mg, about 7175mg, about 7200 mg, about 7225 mg, about 7250 mg, about 7275 mg, about7300 mg, about 7325 mg, about 7350 mg, about 7375 mg, about 7400 mg,about 7425 mg, about 7450 mg, about 7475 mg, about 7500 mg, about 7525mg, about 7550 mg, about 7575 mg, about 7600 mg, about 7625 mg, about7650 mg, about 7675 mg, about 7700 mg, about 7725 mg, about 7750 mg,about 7775 mg, about 7800 mg, about 7825 mg, about 7850 mg, about 7875mg, about 7900 mg, about 7925 mg, about 7950 mg, about 7975 mg, about8000 mg, about 8025 mg, about 8050 mg, about 8075 mg, about 8100 mg,about 8125 mg, about 8150 mg, about 8175 mg, about 8200 mg, about 8225mg, about 8250 mg, about 8275 mg, about 8300 mg, about 8325 mg, about8350 mg, about 8375 mg, about 8400 mg, about 8425 mg, about 8450 mg,about 8475 mg, about 8500 mg, about 8525 mg, about 8550 mg, about 8575mg, about 8600 mg, about 8625 mg, about 8650 mg, about 8675 mg, about8700 mg, about 8725 mg, about 8750 mg, about 8775 mg, about 8800 mg,about 8825 mg, about 8850 mg, about 8875 mg, about 8900 mg, about 8925mg, about 8950 mg, about 8975 mg, about 9000 mg, about 9025 mg, about9050 mg, about 9075 mg, about 9100 mg, about 9125 mg, about 9150 mg,about 9175 mg, about 9200 mg, about 9225 mg, about 9250 mg, about 9275mg, about 9300 mg, about 9325 mg, about 9350 mg, about 9375 mg, about9400 mg, about 9425 mg, about 9450 mg, about 9475 mg, about 9500 mg,about 9525 mg, about 9550 mg, about 9575 mg, about 9600 mg, about 9625mg, about 9650 mg, about 9675 mg, about 9700 mg, about 9725 mg, about9750 mg, about 9775 mg, about 9800 mg, about 9825 mg, about 9850 mg,about 9875 mg, about 9900 mg, about 9925 mg, about 9950 mg, about 9975mg, or about 10,000 mg.

In another embodiment, any of the methods disclosed herein are used intreatment or prevention of a subject or subjects that consume atraditional Western diet. In one embodiment, the methods of theinvention include a step of identifying a subject as a Western dietconsumer or prudent diet consumer and then treating the subject if thesubject is deemed a Western diet consumer. The term “Western diet”herein refers generally to a typical diet consisting of, by percentageof total calories, about 45% to about 50% carbohydrate, about 35% toabout 40% fat, and about 10% to about 15% protein. A Western diet mayalternately or additionally be characterized by relatively high intakesof red and processed meats, sweets, refined grains, and desserts, forexample more than 50%, more than 60% or more or 70% of total caloriescome from these sources.

In one embodiment, a composition for use in methods of the inventioncomprises eicosapentaenoic acid, or a pharmaceutically acceptable ester,derivative, conjugate or salt thereof, or mixtures of any of theforegoing, collectively referred to herein as “EPA.” The term“pharmaceutically acceptable” in the present context means that thesubstance in question does not produce unacceptable toxicity to thesubject or interaction with other components of the composition.

In one embodiment, the EPA comprises all-ciseicosa-5,8,11,14,17-pentaenoic acid. In another embodiment, the EPAcomprises an eicosapentaenoic acid ester. In another embodiment, the EPAcomprises a C₁-C₅ alkyl ester of eicosapentaenoic acid. In anotherembodiment, the EPA comprises eicosapentaenoic acid ethyl ester,eicosapentaenoic acid methyl ester, eicosapentaenoic acid propyl ester,or eicosapentaenoic acid butyl ester. In another embodiment, the EPAcomprises In one embodiment, the EPA comprises all-ciseicosa-5,8,11,14,17-pentaenoic acid ethyl ester.

In another embodiment, the EPA is in the form of ethyl-EPA, lithium EPA,mono-, di- or triglyceride EPA or any other ester or salt of EPA, or thefree acid form of EPA. The EPA may also be in the form of a2-substituted derivative or other derivative which slows down its rateof oxidation but does not otherwise change its biological action to anysubstantial degree.

In another embodiment, EPA is present in a composition useful inaccordance with methods of the invention in an amount of about 50 mg toabout 5000 mg, about 75 mg to about 2500 mg, or about 100 mg to about1000 mg, for example about 75 mg, about 100 mg, about 125 mg, about 150mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400mg, about 425 mg, about 450 mg, about 475 mg, about 500 mg, about 525mg, about 550 mg, about 575 mg, about 600 mg, about 625 mg, about 650mg, about 675 mg, about 700 mg, about 725 mg, about 750 mg, about 775mg, about 800 mg, about 825 mg, about 850 mg, about 875 mg, about 900mg, about 925 mg, about 950 mg, about 975 mg, about 1000 mg, about 1025mg, about 1050 mg, about 1075 mg, about 1100 mg, about 1025 mg, about1050 mg, about 1075 mg, about 1200 mg, about 1225 mg, about 1250 mg,about 1275 mg, about 1300 mg, about 1325 mg, about 1350 mg, about 1375mg, about 1400 mg, about 1425 mg, about 1450 mg, about 1475 mg, about1500 mg, about 1525 mg, about 1550 mg, about 1575 mg, about 1600 mg,about 1625 mg, about 1650 mg, about 1675 mg, about 1700 mg, about 1725mg, about 1750 mg, about 1775 mg, about 1800 mg, about 1825 mg, about1850 mg, about 1875 mg, about 1900 mg, about 1925 mg, about 1950 mg,about 1975 mg, about 2000 mg, about 2025 mg, about 2050 mg, about 2075mg, about 2100 mg, about 2125 mg, about 2150 mg, about 2175 mg, about2200 mg, about 2225 mg, about 2250 mg, about 2275 mg, about 2300 mg,about 2325 mg, about 2350 mg, about 2375 mg, about 2400 mg, about 2425mg, about 2450 mg, about 2475 mg, about 2500 mg, about 2525 mg, about2550 mg, about 2575 mg, about 2600 mg, about 2625 mg, about 2650 mg,about 2675 mg, about 2700 mg, about 2725 mg, about 2750 mg, about 2775mg, about 2800 mg, about 2825 mg, about 2850 mg, about 2875 mg, about2900 mg, about 2925 mg, about 2950 mg, about 2975 mg, about 3000 mg,about 3025 mg, about 3050 mg, about 3075 mg, about 3100 mg, about 3125mg, about 3150 mg, about 3175 mg, about 3200 mg, about 3225 mg, about3250 mg, about 3275 mg, about 3300 mg, about 3325 mg, about 3350 mg,about 3375 mg, about 3400 mg, about 3425 mg, about 3450 mg, about 3475mg, about 3500 mg, about 3525 mg, about 3550 mg, about 3575 mg, about3600 mg, about 3625 mg, about 3650 mg, about 3675 mg, about 3700 mg,about 3725 mg, about 3750 mg, about 3775 mg, about 3800 mg, about 3825mg, about 3850 mg, about 3875 mg, about 3900 mg, about 3925 mg, about3950 mg, about 3975 mg, about 4000 mg, about 4025 mg, about 4050 mg,about 4075 mg, about 4100 mg, about 4125 mg, about 4150 mg, about 4175mg, about 4200 mg, about 4225 mg, about 4250 mg, about 4275 mg, about4300 mg, about 4325 mg, about 4350 mg, about 4375 mg, about 4400 mg,about 4425 mg, about 4450 mg, about 4475 mg, about 4500 mg, about 4525mg, about 4550 mg, about 4575 mg, about 4600 mg, about 4625 mg, about4650 mg, about 4675 mg, about 4700 mg, about 4725 mg, about 4750 mg,about 4775 mg, about 4800 mg, about 4825 mg, about 4850 mg, about 4875mg, about 4900 mg, about 4925 mg, about 4950 mg, about 4975 mg, or about5000 mg.

In another embodiment, a composition useful in accordance with theinvention contains not more than about 10%, not more than about 9%, notmore than about 8%, not more than about 7%, not more than about 6%, notmore than about 5%, not more than about 4%, not more than about 3%, notmore than about 2%, not more than about 1%, or not more than about 0.5%,by weight of all fatty acids (and/or derivatives thereof) present,docosahexaenoic acid (DHA), if any. In another embodiment, a compositionof the invention contains substantially no docosahexaenoic acid. Instill another embodiment, a composition useful in the present inventioncontains no docosahexaenoic acid and/or derivative thereof.

In another embodiment, EPA comprises at least 70%, at least 80%, atleast 90%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100%, by weight of all fatty acids (and/or derivativesthereof) present, in a composition that is useful in methods of thepresent invention.

In one embodiment, a composition of the invention comprises ultra-pureEPA. The term “ultra-pure” as used herein with respect to EPA refers toa composition comprising at least 95%, by weight of all fatty acids(and/or derivatives thereof) present, EPA (as the term “EPA” is definedand exemplified herein). Ultra-pure EPA comprises at least 96%, byweight of all fatty acids (and/or derivatives thereof) present, EPA, atleast 97%, by weight of all fatty acids (and/or derivatives thereof)present, EPA, or at least 98%, by weight of all fatty acids (and/orderivatives thereof) present, EPA, wherein the EPA is any form of EPA asset forth herein.

In another embodiment, a composition useful in accordance with methodsof the invention contains less than 10%, less than 9%, less than 8%,less than 7%, less than 6%, less than 5%, less than 4%, less than 3%,less than 2%, less than 1%, less than 0.5% or less than 0.25%, by weightof all fatty acids (and/or derivatives thereof) present, of any fattyacid other than EPA. Illustrative examples of a “fatty acid other thanEPA” include linolenic acid (LA), arachidonic acid (AA), docosahexaenoicacid (DHA), alpha-linolenic acid (ALA), stearadonic acid (STA),eicosatrienoic acid (ETA) and/or docosapentaenoic acid (DPA). In anotherembodiment, a composition useful in accordance with methods of theinvention contains about 0.1% to about 4%, about 0.5% to about 3%, orabout 1% to about 2%, by weight of all fatty acids (and/or derivativesthereof) present, other than EPA and/or DHA.

In another embodiment, a composition useful in accordance with theinvention has one or more of the following features: (a)eicosapentaenoic acid ethyl ester represents at least about 96%, atleast about 97%, or at least about 98%, by weight of all fatty acids(and/or derivatives thereof) present, in the composition; (b) thecomposition contains not more than about 4%, not more than about 3%, ornot more than about 2%, by weight of all fatty acids (and/or derivativesthereof) present, other than eicosapentaenoic acid ethyl ester; (c) thecomposition contains not more than about 0.6%, not more than about 0.5%,or not more than about 0.4%, by weight of all fatty acids (and/orderivatives thereof) present, of any individual fatty acid other thaneicosapentaenoic acid ethyl ester; (d) the composition has a refractiveindex (20° C.) of about 1 to about 2, about 1.2 to about 1.8 or about1.4 to about 1.5; (e) the composition has a specific gravity (20° C.) ofabout 0.8 to about 1.0, about 0.85 to about 0.95 or about 0.9 to about0.92; (e) the composition contains not more than about 20 ppm, not morethan about 15 ppm or not more than about 10 ppm heavy metals, (f) thecomposition contains not more than about 5 ppm, not more than about 4ppm, not more than about 3 ppm, or not more than about 2 ppm arsenic,and/or (g) the composition has a peroxide value of not more than about 5meq/kg, not more than about 4 meq/kg, not more than about 3 meq/kg, ornot more than about 2 meq/kg.

In another embodiment, a composition useful in accordance with theinvention comprises, consists of or consists essentially of at least95%, by weight of all fatty acids (and/or derivatives thereof) present,ethyl eicosapentaenoate (EPA-E), about 0.2% to about 0.5%, by weight ofall fatty acids (and/or derivatives thereof) present, ethyloctadecatetraenoate (ODTA-E), about 0.05% to about 0.25%, by weight ofall fatty acids (and/or derivatives thereof) present, ethylnonadecapentaenoate (NDPA-E), about 0.2% to about 0.45%, by weight ofall fatty acids (and/or derivatives thereof) present, ethyl arachidonate(AA-E), about 0.3% to about 0.5%, by weight of all fatty acids (and/orderivatives thereof) present, ethyl eicosatetraenoate (ETA-E), and about0.05% to about 0.32%, by weight of all fatty acids (and/or derivativesthereof) present, ethyl heneicosapentaenoate (HPA-E). In anotherembodiment, the composition is present in a capsule shell.

In another embodiment, compositions useful in accordance with theinvention comprise, consist essential of, or consist of at least 95%,96% or 97%, by weight of all fatty acids (and/or derivatives thereof)present, ethyl eicosapentaenoate, about 0.2% to about 0.5% by weightethyl octadecatetraenoate, about 0.05% to about 0.25%, by weight of allfatty acids (and/or derivatives thereof) present, ethylnonadecapentaenoate, about 0.2% to about 0.45%, by weight of all fattyacids (and/or derivatives thereof) present, ethyl arachidonate, about0.3% to about 0.5%, by weight of all fatty acids (and/or derivativesthereof) present, ethyl eicosatetraenoate, and about 0.05% to about0.32%, by weight of all fatty acids (and/or derivatives thereof)present, ethyl heneicosapentaenoate. Optionally, the compositioncontains not more than about 0.06%, about 0.05%, or about 0.04%, byweight of all fatty acids (and/or derivatives thereof) present, DHA orderivative thereof such as ethyl-DHA. In one embodiment the compositioncontains substantially no or no amount of DHA or derivative thereof suchas ethyl-DHA. The composition further optionally comprises one or moreantioxidants (e.g. tocopherol) or other impurities in an amount of notmore than about 0.5% or not more than 0.05%. In another embodiment, thecomposition comprises about 0.05% to about 0.4%, for example about 0.2%by weight tocopherol. In another embodiment, about 500 mg to about 1 gof the composition is provided in a capsule shell.

In another embodiment, compositions useful in accordance with theinvention comprise, consist essential of, or consist of at least 96%, byweight of all fatty acids (and/or derivatives thereof) present, ethyleicosapentaenoate, about 0.22% to about 0.4%, by weight of all fattyacids (and/or derivatives thereof) present, ethyl octadecatetraenoate,about 0.075% to about 0.20%, by weight of all fatty acids (and/orderivatives thereof) present, ethyl nonadecapentaenoate, about 0.25% toabout 0.40%, by weight of all fatty acids (and/or derivatives thereof)present, ethyl arachidonate, about 0.3% to about 0.4%, by weight of allfatty acids (and/or derivatives thereof) present, ethyleicosatetraenoate and about 0.075% to about 0.25%, by weight of allfatty acids (and/or derivatives thereof) present, ethylheneicosapentaenoate. Optionally, the composition contains not more thanabout 0.06%, about 0.05%, or about 0.04%, by weight of all fatty acids(and/or derivatives thereof) present, DHA or derivative thereof such asethyl-DHA. In one embodiment the composition contains substantially noor no amount of DHA or derivative thereof such as ethyl-DHA. Thecomposition further optionally comprises one or more antioxidants (e.g.tocopherol) or other impurities in an amount of not more than about 0.5%or not more than 0.05%. In another embodiment, the composition comprisesabout 0.05% to about 0.4%, for example about 0.2% by weight tocopherol.In another embodiment, the invention provides a dosage form comprisingabout 500 mg to about 1 g of the foregoing composition in a capsuleshell. In one embodiment, the dosage form is a gel or liquid capsule andis packaged in blister packages of about 1 to about 20 capsules persheet.

In another embodiment, compositions useful in accordance with theinvention comprise, consist essential of, or consist of at least 96%,97% or 98%, by weight of all fatty acids (and/or derivatives thereof)present, ethyl eicosapentaenoate, about 0.25% to about 0.38%, by weightof all fatty acids (and/or derivatives thereof) present, ethyloctadecatetraenoate, about 0.10% to about 0.15%, by weight of all fattyacids (and/or derivatives thereof) present, ethyl nonadecapentaenoate,about 0.25% to about 0.35%, by weight of all fatty acids (and/orderivatives thereof) present, ethyl arachidonate, about 0.31% to about0.38%, by weight of all fatty acids (and/or derivatives thereof)present, ethyl eicosatetraenoate, and about 0.08% to about 0.20%, byweight of all fatty acids (and/or derivatives thereof) present, ethylheneicosapentaenoate. Optionally, the composition contains not more thanabout 0.06%, about 0.05%, or about 0.04%, by weight of all fatty acids(and/or derivatives thereof) present, DHA or derivative thereof such asethyl-DHA. In one embodiment the composition contains substantially noor no amount of DHA or derivative thereof such as ethyl-DHA. Thecomposition further optionally comprises one or more antioxidants (e.g.tocopherol) or other impurities in an amount of not more than about 0.5%or not more than 0.05%. In another embodiment, the composition comprisesabout 0.05% to about 0.4%, for example about 0.2% by weight tocopherol.In another embodiment, the invention provides a dosage form comprisingabout 500 mg to about 1 g of the foregoing composition in a capsuleshell.

In another embodiment, a composition as described herein is administeredto a subject once or twice per day. In another embodiment, 1, 2, 3 or 4capsules, each containing about 1 g of a composition as describedherein, are administered to a subject daily. In another embodiment, 1 or2 capsules, each containing about 1 g of a composition as describedherein, are administered to the subject in the morning, for examplebetween about 5 am and about 11 am, and 1 or 2 capsules, each containingabout 1 g of a composition as described herein, are administered to thesubject in the evening, for example between about 5 pm and about 11 pm.

In one embodiment, a subject being treated in accordance with methods ofthe invention is not otherwise on lipid-altering therapy, for examplestatin, fibrate, niacin and/or ezetimibe therapy.

In another embodiment, compositions useful in accordance with methods ofthe invention are orally deliverable. The terms “orally deliverable” or“oral administration” herein include any form of delivery of atherapeutic agent or a composition thereof to a subject wherein theagent or composition is placed in the mouth of the subject, whether ornot the agent or composition is swallowed. Thus “oral administration”includes buccal and sublingual as well as esophageal administration. Inone embodiment, the composition is present in a capsule, for example asoft gelatin capsule.

In some embodiments, a composition according to the present inventioncomprises rosiglitazone and provides a mean steady state plasma C_(max),a mean steady state plasma AUC_(0-inf), and/or a mean steady stateplasma T_(max) of rosiglitazone of about 70% to about 135%, whenco-administered with about 2 g or about 4 g per day of ethyleicosapentaenoate, of a mean steady state plasma C_(max), a mean steadystate plasma AUC_(0-inf), and/or a mean steady state plasma T_(max) ofrosiglitazone provided by a second composition comprising rosiglitazoneadministered without the ethyl eicosapentaenoate. In some embodiments,the composition provides a mean steady state plasma C_(max), a meansteady state plasma AUC_(0-inf), and/or a mean steady state plasmaT_(max) of rosiglitazone of about 80% to about 125% of a mean steadystate plasma C_(max), a mean steady state plasma AUC_(0-inf), and/or amean steady state plasma T_(max) of rosiglitazone provided by the secondcomposition. In some embodiments, the composition provides a mean steadystate plasma C_(max), a mean steady state plasma AUC_(0-inf), and/or amean steady state plasma T_(max) of rosiglitazone of about 70% to about135%, when co-administered with about 4 g per day of ethyleicosapentaenoate, compared to a mean steady state plasma C_(max), amean steady state plasma AUC_(0-inf), and/or a mean steady state plasmaT_(max) of rosiglitazone provided by the second composition. In someembodiments, the composition provides a mean steady state AUC_(0-inf) ofabout 80% to about 125% of the second composition. In some embodiments,the composition provides a mean steady state AUC_(0-inf) of about 168.6ng·hr/mL. In some embodiments, the composition provides a mean steadystate C_(max) of about 70% to about 135%, when co-administered withabout 4 g per day of ethyl eicosapentaenoate, of a mean steady stateplasma C_(max) provided by the second composition. In some embodiments,the composition provides a mean steady state C_(max) of about 80% toabout 125% of the second composition. In some embodiments, thecomposition provides a mean steady state C_(max) of about 53.2 ng/mL.

In any embodiment disclosed herein, rosiglitazone may be present in anamount of about 1 mg to about 160 mg, for example about 1 mg, about 2mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8mg, about 9 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg,about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg,about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about110 mg, about 115 mg, about 120 mg, about 125 mg, about 130 mg, about135 mg, about 140 mg, about 145 mg, about 150 mg, about 155 mg, or about160 mg.

In some embodiments, a composition of the present invention comprises atleast about 80%, by weight of all fatty acids (and/or derivativesthereof) present, ethyl eicosapentaenoate, wherein the composition doesnot significantly alter a blood plasma C_(max), a blood plasmaAUC_(0-inf), and/or a blood plasma T_(max) of rosiglitazone. In someembodiments, the composition is administered at a daily dose of about 2g or about 4 g per day. In some embodiments, the rosiglitazone isadministered at a daily dose of about 80 mg per day. In someembodiments, the blood plasma C_(max), the blood plasma AUC_(0-inf),and/or the blood plasma T_(max) is a steady state blood plasma C_(max),a steady state blood plasma AUC_(0-inf), and/or a steady state bloodplasma T_(max). In some embodiments, the composition alters the bloodplasma C_(max), the blood plasma AUC_(0-inf), and/or the blood plasmaT_(max) of rosiglitazone by no more than about 30%, by no more thanabout 25%, by no more than about 20%, or by no more than about 15%compared to administration of rosiglitazone without the composition. Insome embodiments, the composition alters the blood plasma C_(max) andthe blood plasma AUC_(0-inf) of rosiglitazone by no more than about 35%,by no more than about 30%, by no more than about 25%, by no more thanabout 20%, or by no more than about 15% compared to administration ofrosiglitazone without the composition. In some embodiments, thecomposition alters the blood plasma T_(max) and the blood plasmaAUC_(0-inf) of rosiglitazone by no more than about 35%, by no more thanabout 30%, by no more than about 25%, by no more than about 20%, or byno more than about 15% compared to administration of rosiglitazonewithout the composition. In some embodiments, the composition alters theblood plasma C_(max) and the blood plasma T_(max) of rosiglitazone by nomore than about 35%, by no more than about 30%, by no more than about25%, by no more than about 20%, or by no more than about 15% compared toadministration of rosiglitazone without the composition. In someembodiments, the composition alters the blood plasma C_(max), the bloodplasma AUC_(0-inf), and the blood plasma T_(max) of rosiglitazone by nomore than about 35%, by no more than about 30%, by no more than about25%, by no more than about 20%, or by no more than about 15% compared toadministration of rosiglitazone without the composition.

In some embodiments, a method of reducing triglycerides in a subject onrosiglitazone therapy according to the present invention comprisesadministering to the subject a composition comprising at least about80%, by weight of all fatty acids (and/or derivatives thereof) present,ethyl eicosapentaenoate. In other embodiments, a method of reducingtriglycerides in a subject on rosiglitazone therapy according to thepresent invention comprises administering to the subject about 1 toabout 4 capsules per day, each capsule comprising about 1 g of ethyleicosapentaenoate. In some embodiments, the capsules comprise at leastabout 80%, by weight of all fatty acids (and/or derivatives thereof)present, ethyl eicosapentaenoate. In some embodiments, a C_(max), anAUC_(0-inf), and/or a T_(max) of rosiglitazone is not significantlyaltered compared to a second subject or a second subject group who hasreceived the rosiglitazone but not the ethyl eicosapentaenoate. In someembodiments, any one or more of the C_(max), the AUC_(0-inf), and/or theT_(max) of rosiglitazone is altered by no more than about 35%, by nomore than about 30%, by no more than about 25%, by no more than about20%, or by no more than about 15% compared to the second subject orsecond subject group. In some embodiments, the subject has a fastingbaseline triglyceride level of about 200 mg/dl to 499 mg/dl. In someembodiments, the second subject or second subject group has a fastingbaseline triglyceride level or a mean or median fasting baselinetriglyceride level of about 200 mg/dl to 499 mg/dl. In some embodiments,the subject has a fasting baseline triglyceride level of at least 500mg/dl. In some embodiments, the second subject or second subject grouphas a fasting baseline triglyceride level or a mean or median fastingbaseline triglyceride level of at least 500 mg/dl. In some embodiments,triglycerides are reduced in the subject with no increase in an LDL-Clevel in the subject. In some embodiments, the reduction intriglycerides and the no increase in LDL-C level is in comparison tobaseline or to a second subject or subject group that has receivedrosiglitazone but not the ethyl eicosapentaenoate.

A composition for use in accordance with the invention can be formulatedas one or more dosage units. The terms “dose unit” and “dosage unit”herein refer to a portion of a pharmaceutical composition that containsan amount of a therapeutic agent suitable for a single administration toprovide a therapeutic effect. Such dosage units may be administered oneto a plurality (i.e. 1 to about 10, 1 to 8, 1 to 6, 1 to 4 or 1 to 2) oftimes per day, or as many times as needed to elicit a therapeuticresponse.

In another embodiment, the invention provides use of any compositiondescribed herein for treating moderate to severe hypertriglyceridemia ina subject in need thereof, comprising: providing a subject having afasting baseline triglyceride level of 500 mg/dl to about 1500 mg/dl andadministering to the subject a pharmaceutical composition as describedherein. In one embodiment, the composition comprises about 1 g to about4 g of eicosapentaenoic acid ethyl ester, wherein the compositioncontains substantially no docosahexaenoic acid. In some embodiments,cholesterol domain formation in membranes of the subject is reduced orprevented. In some embodiments, the subject experiences no substantialincrease, or no increase, or a reduction, in LDL-C levels.

In another embodiment, the invention provides use of any compositiondescribed herein for treating moderate to severe hypertriglyceridemia ina subject in need thereof, comprising: providing a subject on statintherapy and having a fasting baseline triglyceride level of about 200mg/dl to 499 mg/dl and administering to the subject a pharmaceuticalcomposition as described herein. In one embodiment, the compositioncomprises about 1 g to about 4 g of eicosapentaenoic acid ethyl ester,wherein the composition contains substantially no docosahexaenoic acid.In some embodiments, cholesterol domain formation in membranes of thesubject is reduced or prevented. In some embodiments, the subjectexperiences no substantial increase, or no increase, or a reduction, inLDL-C levels.

In one embodiment, compositions of the invention, upon storage in aclosed container maintained at room temperature, refrigerated (e.g.about 5 to about 5-10° C.) temperature, or frozen for a period of about1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, exhibit at least about90%, at least about 95%, at least about 97.5%, or at least about 99% ofthe active ingredient(s) originally present therein.

In one embodiment, the invention provides use of a composition asdescribed herein in manufacture of a medicament for treatment of any ofa cardiovascular-related disease. In another embodiment, the subject isdiabetic.

In one embodiment, a composition as set forth herein is packagedtogether with instructions for using the composition to treat acardiovascular disorder.

EXAMPLES Example 1

A multi-center, placebo-controlled randomized, double-blind, 12-weekstudy with an open-label extension was performed to evaluate theefficacy and safety of AMR101 in patients with fasting triglyceridelevels ≧500 mg/dL. The primary objective of the study was to determinethe efficacy of AMR101 2 g daily and 4 g daily, compared to placebo, inlowering fasting TG levels in patients with fasting TG levels 500 mg/dLand 1500 mg/dL (5.65 mmol/L and 16.94 mmol/L).

The secondary objectives of this study were the following:

-   1. To determine the safety and tolerability of AMR101 2 g daily and    4 g daily;-   2. To determine the effect of AMR101 on lipid and apolipoprotein    profiles;-   3. To determine the effect of AMR101 on low-density lipoprotein    (LDL) particle number and size;-   4. To determine the effect of AMR101 on oxidized LDL;-   5. To determine the effect of AMR101 on fasting plasma glucose (FPG)    and hemoglobin A_(lc) (HbA_(lc));-   6. To determine the effect of AMR101 on insulin resistance;-   7. To determine the effect of AMR101 on high-sensitivity C-reactive    protein (hsCRP);-   8. To determine the effects of AMR101 2 g daily and 4 g daily on the    incorporation of fatty acids into red blood cell membranes and into    plasma phospholipids;-   9. To explore the relationship between baseline fasting TG levels    and the reduction in fasting TG levels; and-   10. To explore the relationship between an increase in red blood    cell membrane eicosapentaenoic acid (EPA) concentrations and the    reduction in fasting TG levels.

The population for this study was men and women (women of childbearingpotential needed to be on contraception or practice abstinence)>18 yearsof age with a body mass index ≦45 kg/m² who were not on lipid-alteringtherapy or were not currently on lipid-altering therapy. Patientscurrently on statin therapy (with or without ezetimibe) were evaluatedby the investigator as to whether this therapy could be safelydiscontinued at screening, or if it should have been continued. Ifstatin therapy (with or without ezetimibe) was to be continued, dose(s)must have been stable for ≧4 weeks prior to randomization. Patientstaking non-statin, lipid-altering medications (niacin >200 mg/day,fibrates, fish oil, other products containing omega-3 fatty acids, orother herbal products or dietary supplements with potentiallipid-altering effects), either alone or in combination with statintherapy (with or without ezetimibe), must have been able to safelydiscontinue non-statin, lipid-altering therapy at screening.

Approximately 240 patients were randomized at approximately 50 centersin North America, South America, Central America, Europe, India, andSouth Africa. The study was a 58- to 60-week, Phase 3, multi-centerstudy consisting of 3 study periods: (1) a 6- to 8-week screening periodthat included a diet and lifestyle stabilization and washout period anda TG qualifying period; (2) a 12-week, double-blind, randomized,placebo-controlled treatment period; and (3) a 40-week, open-label,extension period.

During the screening period and double-blind treatment period, allvisits were within ±3 days of the scheduled time. During the open-labelextension period, all visits were within ±7 days of the scheduled time.The screening period included a 4- or 6-week diet and lifestylestabilization period and washout period followed by a 2-week TGqualifying period.

The screening visit (Visit 1) occurred for all patients at either 6weeks (for patients not on lipid-altering therapy at screening or forpatients who did not need to discontinue their current lipid-alteringtherapy) or 8 weeks (for patients who required washout of their currentlipid-altering therapy at screening) before randomization, as follows:

Patients who did not require a washout: The screening visit will occurat Visit 1 (Week-6). Eligible patients entered a 4-week diet andlifestyle stabilization period. At the screening visit, all patientsreceived counseling regarding the importance of the National CholesterolEducation Program (NCEP) Therapeutic Lifestyle Changes (TLC) diet andreceived instructions on how to follow this diet. Patients who requireda washout: The screening visit occurred at Visit 1 (Week-8). Eligiblepatients began a 6-week washout period at the screening visit. Patientsreceived counseling regarding the NCEP TLC diet and receivedinstructions on how to follow this diet. Site personnel contactedpatients who did not qualify for participation based on screeninglaboratory test results to instruct them to resume their priorlipid-altering medications.

At the end of the 4-week diet and lifestyle stabilization period or the6-week diet and stabilization and washout period, eligible patientsentered the 2-week TG qualifying period and had their fasting TG levelmeasured at Visit 2 (Week-2) and Visit 3 (Week-1). Eligible patientsmust have had an average fasting TG level ≧500 mg/dL and ≦1500 mg/dL(≧5.65 mmol/L and ≦16.94 mmol/L) to enter the 12-week double-blindtreatment period. The TG level for qualification was based on theaverage (arithmetic mean) of the Visit 2 (Week-2) and Visit 3 (Week-1)values. If a patient's average TG level from Visit 2 and Visit 3 felloutside the required range for entry into the study, an additionalsample for fasting TG measurement was collected 1 week later at Visit3.1. If a third sample was collected at Visit 3.1, entry into the studywas based on the average (arithmetic mean) of the values from Visit 3and Visit 3.1.

After confirmation of qualifying fasting TG values, eligible patientsentered a 12-week, randomized, double-blind treatment period. At Visit 4(Week 0), patients were randomly assigned to one of the followingtreatment groups:

-   -   AMR101 2 g daily,    -   AMR101 4 g daily, or    -   Placebo.

During the double-blind treatment period, patients returned to the siteat Visit 5 (Week 4), Visit 6 (Week 11), and Visit 7 (Week 12) forefficacy and safety evaluations.

Patients who completed the 12-week double-blind treatment period wereeligible to enter a 40-week, open-label, extension period at Visit 7(Week 12). All patients received open-label AMR101 4 g daily. From Visit8 (Week 16) until the end of the study, changes to the lipid-alteringregimen were permitted (e.g., initiating or raising the dose of statinor adding non-statin, lipid-altering medications to the regimen), asguided by standard practice and prescribing information. After Visit 8(Week 16), patients returned to the site every 12 weeks until the lastvisit at Visit 11 (Week 52).

Eligible patients were randomly assigned at Visit 4 (Week 0) to orallyreceive AMR101 2 g daily, AMR101 4 g daily, or placebo for the 12-weekdouble-blind treatment period. AMR101 was provided in 1 g liquid-filled,oblong, gelatin capsules. The matching placebo capsule was filled withlight liquid paraffin and contained 0 g of AMR101. During thedouble-blind treatment period, patients took 2 capsules (AMR101 ormatching placebo) in the morning and 2 in the evening for a total of 4capsules per day. Patients in the AMR101 2 g/day treatment groupreceived 1 AMR101 1 g capsule and 1 matching placebo capsule in themorning and in the evening. Patients in the AMR101 4 g/day treatmentgroup received 2 AMR101 1 g capsules in the morning and evening.

Patients in the placebo group received 2 matching placebo capsules inthe morning and evening. During the extension period, patients receivedopen-label AMR101 4 g daily. Patients took 2 AMR101 1 g capsules in themorning and 2 in the evening.

The primary efficacy variable for the double-blind treatment period waspercent change in TG from baseline to Week 12 endpoint. The secondaryefficacy variables for the double-blind treatment period included thefollowing:

-   -   Percent changes in total cholesterol (TC), high-density        lipoprotein cholesterol (HDL-C), calculated low-density        lipoprotein cholesterol (LDL-C), calculated non-high-density        lipoprotein cholesterol (non-HDL-C), and very low-density        lipoprotein cholesterol (VLDL-C) from baseline to Week 12        endpoint;    -   Percent change in very low-density lipoprotein TG from baseline        to Week 12;    -   Percent changes in apolipoprotein A-I (apo A-I), apolipoprotein        B (apo B), and apo A-I/apo B ratio from baseline to Week 12;    -   Percent changes in lipoprotein(a) from baseline to Week 12        (selected sites only);    -   Percent changes in LDL particle number and size, measured by        nuclear magnetic resonance, from baseline to Week 12 (selected        sites only);    -   Percent change in remnant-like particle cholesterol from        baseline to Week 12 (selected sites only);    -   Percent change in oxidized LDL from baseline to Week 12        (selected sites only);    -   Changes in FPG and HbA_(lc) from baseline to Week 12;    -   Change in insulin resistance, as assessed by the homeostasis        model index insulin resistance, from baseline to Week 12;    -   Percent change in lipoprotein associated phospholipase A2 from        baseline to Week 12 (selected sites only);    -   Change in intracellular adhesion molecule-1 from baseline to        Week 12 (selected sites only);    -   Change in interleukin-6 from baseline to Week 12 (selected sites        only);    -   Change in plasminogen activator inhibitor-1 from baseline to        Week 12 (selected sites only);    -   Change in hsCRP from baseline to Week 12 (selected sites only);    -   Change in serum phospholipid EPA content from baseline to Week        12;    -   Change in red blood cell membrane EPA content from baseline to        Week 12; and    -   Change in serum phospholipid and red blood cell membrane content        in the following fatty acids from baseline to Week 12:        docosapentaenoic acid, docosahexaenoic acid, arachidonic acid,        palmitic acid, stearic acid, and oleic acid.

The efficacy variable for the open-label extension period was percentchange in fasting TG from extension baseline to end of treatment. Safetyassessments included adverse events, clinical laboratory measurements(chemistry, hematology, and urinalysis), 12-lead electrocardiograms(ECGs), vital signs, and physical examinations

For TG, TC, HDL-C, calculated LDL-C, calculated non-HDL-C, and VLDL-C,baseline was defined as the average of Visit 4 (Week 0) and thepreceding lipid qualifying visit (either Visit 3 [Week-1] or if itoccurs, Visit 3.1) measurements. Baseline for all other efficacyparameters was the Visit 4 (Week 0) measurement.

For TC, HDL-C, calculated LDL-C, calculated non-HDL-C, and VLDL-C, Week12 endpoint was defined as the average of Visit 6 (Week 11) and Visit 7(Week 12) measurements. Week 12 endpoint for all other efficacyparameters was the Visit 7 (Week 12) measurement.

The primary efficacy analysis was performed using a 2-way analysis ofcovariance (ANCOVA) model with treatment as a factor and baseline TGvalue as a covariate. The least-squares mean, standard error, and2-tailed 95% confidence interval for each treatment group and for eachcomparison was estimated. The same 2-way ANCOVA model was used for theanalysis of secondary efficacy variables.

The primary analysis was repeated for the per-protocol population toconfirm the robustness of the results for the intent-to-treatpopulation.

The primary efficacy variable was the percent change in fasting TGlevels from baseline to Week 12. A sample size of 69 completed patientsper treatment group was expected to provide ≧90% power to detect adifference of 30% between AMR101 and placebo in percent change frombaseline in fasting TG levels, assuming a standard deviation of 45% inTG measurements and a significance level of p<0.01. To accommodate a 15%drop-out rate from randomization to completion of the double-blindtreatment period, a total of 240 randomized patients was planned (80patients per treatment group).

Example 2

A multi-center, placebo-controlled, randomized, double-blind, 12-weekstudy was performed to evaluate the efficacy and safety of >96% E-EPA inpatients with fasting triglyceride levels ≧200 mg/dl and <500 mg/dldespite statin therapy (the mean of two qualifying entry values neededto be ≧185 mg/dl and at least one of the values needed to be ≧200mg/dl). The primary objective of the study was to determine the efficacyof >96% E-EPA 2 g daily and 4 g daily, compared to placebo, in loweringfasting TG levels in patients with high risk for cardiovascular diseaseand with fasting TG levels ≧200 mg/dl and <500 mg/dl, despite treatmentto LDL-C goal on statin therapy.

The secondary objectives of this study were the following:

-   -   1. To determine the safety and tolerability of >96% E-EPA 2 g        daily and 4 g daily;    -   2. To determine the effect of >96% E-EPA on lipid and        apolipoprotein profiles including total cholesterol (TC),        non-high-density lipoprotein cholesterol (non-HDL-C), low        density lipoprotein cholesterol (LDL-C), high density        lipoprotein cholesterol (HDL-C), and very high density        lipoprotein cholesterol (VHDL-C);    -   3. To determine the effect of >96% E-EPA on lipoprotein        associated phospholipase A₂ (Lp-PLA₂) from baseline to week 12;    -   4. To determine the effect of >96% E-EPA on low-density        lipoprotein (LDL) particle number and size;    -   5. To determine the effect of >96% E-EPA on oxidized LDL;    -   6. To determine the effect of >96% E-EPA on fasting plasma        glucose (FPG) and hemoglobin A_(lc) (HbA_(lc));    -   7. To determine the effect of >96% E-EPA on insulin resistance;    -   8. To determine the effect of >96% E-EPA on high-sensitivity        C-reactive protein (hsCRP);    -   9. To determine the effects of >96% E-EPA 2 g daily and 4 g        daily on the incorporation of fatty acids into red blood cell        membranes and into plasma phospholipids;    -   10. To explore the relationship between baseline fasting TG        levels and the reduction in fasting TG levels; and    -   11. To explore the relationship between changes of fatty acid        concentrations in plasma and red blood cell membranes, and the        reduction in fasting TG levels.

The population for this study was men and women >18 years of age with abody mass index ≦45 kg/m² with fasting TG levels greater than or equalto 200 mg/dl and less than 500 mg/dl and on a stable does of statintherapy (with or without ezetimibe). The statin was atorvostatin,rosuvastatin or simvastatin. The dose of statin must have been stablefor ≧4 weeks prior to the LDL-C/TG baseline qualifying measurement forrandomization. The statin dose was optimized such that the patients areat their LDL-C goal at the LDL-C/TG baseline qualifying measurements.The same statin at the same dose was continued until the study ended.

Patients taking any additional non-statin, lipid-altering medications(niacin >200 mg/day, fibrates, fish oil, other products containingomega-3 fatty acids, or other herbal products or dietary supplementswith potential lipid-altering effects), either alone or in combinationwith statin therapy (with or without ezetimibe), must have been able tosafely discontinue non-statin, lipid-altering therapy at screening.

Patients at high risk for CVD, i.e., patients with clinical coronaryheart disease (CHD) or clinical CHD risk equivalents (10-year risk >20%)as defined in the National Cholesterol Education Program (NCEP) AdultTreatment Panel III (ATP III)

Guidelines were eligible to participate in this study. Those includedpatients with any of the following criteria: (1) Known CVD, eitherclinical coronary heart disease (CHD), symptomatic carotid arterydisease (CAD), peripheral artery disease (PAD) or abdominal aorticaneurism; or (2) Diabetes Mellitus (Type 1 or 2).

Approximately 702 patients were randomized at approximately 80 centersin the U.S. The study was a 18- to 20-week, Phase 3, multi-center studyconsisting of 2 study periods: (1) A 6- to 8-week screening period thatincluded a diet and lifestyle stabilization, a non-statin lipid-alteringtreatment washout, and an LDL-C and TG qualifying period and (2) A12-week, double-blind, randomized, placebo-controlled treatment period.

During the screening period and double-blind treatment period, allvisits were within ±3 days of the scheduled time. All patients continuedto take the statin product (with or without ezetimibe) at the same dosethey were taking at screening throughout their participation in thestudy.

The 6- to 8-week screening period included a diet and lifestylestabilization, a non-statin lipid-altering treatment washout, and anLDL-C and TG qualifying period. The screening visit (Visit 1) occurredfor all patients at either 6 weeks (for patients on stable statintherapy [with or without ezetimibe] at screening) or 8 weeks (forpatients who will require washout of their current non-statinlipid-altering therapy at screening) before randomization, as follows:

-   -   Patients who did not require a washout: The screening visit        occurred at Visit 1 (Week-6). Eligible patients entered a 4-week        diet and lifestyle stabilization period. At the screening visit,        all patients received counseling regarding the importance of the        National Cholesterol Education Program (NCEP) Therapeutic        Lifestyle Changes (TLC) diet and received basic instructions on        how to follow this diet.    -   Patients who required a washout: The screening visit occurred at        Visit 1 (Week-8). Eligible patients began a 6-week washout        period at the screening visit (i.e. 6 weeks washout before the        first LDL-C/TG qualifying visit). Patients received counseling        regarding the NCEP TLC diet and received basic instructions on        how to follow this diet. Site personnel contacted patients who        did not qualify for participation based on screening laboratory        test results to instruct them to resume their prior        lipid-altering medications.

At the end of the 4-week diet and lifestyle stabilization period or the6-week diet and stabilization and washout period, eligible patientsentered the 2-week LDL-C and TG qualifying period and had their fastingLDL-C and TG levels measured at Visit 2 (Week-2) and Visit 3 (Week-1).Eligible patients must have had an average fasting LDL-C level ≧40 mg/dLand <100 mg/dL and an average fasting TG level ≧200 mg/dL and <500 mg/dLto enter the 12-week double-blind treatment period. The LDL-C and TGlevels for qualification were based on the average (arithmetic mean) ofthe Visit 2 (Week-2) and Visit 3 (Week-1) values. If a patient's averageLDL-C and/or TG levels from Visit 2 and Visit 3 fell outside therequired range for entry into the study, an additional fasting lipidprofile was collected 1 week later at Visit 3.1. If a third sample wascollected at Visit 3.1, entry into the study was based on the average(arithmetic mean) of the values from Visit 3 and Visit 3.1.

After confirmation of qualifying fasting LDL-C and TG values, eligiblepatients entered a 12-week, randomized, double-blind treatment period.At Visit 4 (Week 0), patients were randomly assigned to 1 of thefollowing treatment groups:

-   -   >96% E-EPA 2 g daily,    -   >96% E-EPA 4 g daily, or    -   Placebo.

226 to 234 patients per treatment group were randomized in this study.Stratification was by type of statin (atorvastatin, rosuvastatin orsimvastatin), the presence of diabetes, and gender.

During the double-blind treatment period, patients returned to the siteat Visit 5 (Week 4), Visit 6 (Week 11), and Visit 7 (Week 12) forefficacy and safety evaluations.

Eligible patients were randomly assigned at Visit 4 (Week 0) to receiveorally >96% E-EPA 2 g daily, >96% E-EPA 4 g daily, or placebo.

>96% E-EPA was provided in 1 g liquid-filled, oblong, gelatin capsules.The matching placebo capsule was filled with light liquid paraffin andcontained 0 g of >96% E-EPA. >96% E-EPA capsules were to be taken withfood (i.e. with or at the end of a meal).

During the double-blind treatment period, patients were to take 2capsules (>96% E-EPA or matching placebo) in the morning and 2 capsulesin the evening for a total of 4 capsules per day.

-   -   Patients in the >96% E-EPA 2 g/day treatment group received        1>96% E-EPA 1 g capsule and 1 matching placebo capsule in the        morning and in the evening.    -   Patients in the >96% E-EPA 4 g/day treatment group received        2>96% E-EPA 1 g capsules in the morning and evening.

Patients in the placebo group received 2 matching placebo capsules inthe morning and evening.

The primary efficacy variable for the double-blind treatment period waspercent change in TG from baseline to Week 12 endpoint. The secondaryefficacy variables for the double-blind treatment period included thefollowing:

-   -   Percent changes in total cholesterol (TC), high-density        lipoprotein cholesterol (HDL-C), LDL-C, calculated non-HDL-C,        and very low-density lipoprotein cholesterol (VLDL-C) from        baseline to Week 12 endpoint;    -   Percent change in very low-density lipoprotein TG from baseline        to Week 12;    -   Percent changes in apolipoprotein A-I (apo A-I), apolipoprotein        B (apo B), and apo A-I/apo B ratio from baseline to Week 12;    -   Percent changes in lipoprotein(a) from baseline to Week 12;    -   Percent changes in LDL particle number and size, measured by        nuclear magnetic resonance, from baseline to Week 12;    -   Percent change in remnant-like particle cholesterol from        baseline to Week 12;    -   Percent change in oxidized LDL from baseline to Week 12;    -   Changes in FPG and HbA_(lc) from baseline to Week 12;    -   Change in insulin resistance, as assessed by the homeostasis        model index insulin resistance, from baseline to Week 12;    -   Percent change in lipoprotein associated phospholipase A₂        (Lp-PLA₂) from baseline to Week 12;    -   Change in intracellular adhesion molecule-1 from baseline to        Week 12;    -   Change in interleukin-2 from baseline to Week 12;    -   Change in plasminogen activator inhibitor-1 from baseline to        Week 12. Note: this parameter will only be collected at sites        with proper storage conditions;    -   Change in hsCRP from baseline to Week 12; and    -   Change in plasma concentration and red blood cell membrane        content of fatty acid from baseline to Week 12 including EPA,        docosapentaenoic acid (DPA), docosahexaenoic acid (DHA),        arachidonic acid (AA), dihomo-γ-linolenic acid (DGLA), the ratio        of EPA/AA, ratio of oleic acid/stearic acid (OA/SA), and the        ratio of total omega-3 acids over total omega-6 acids.

Safety assessments included adverse events, clinical laboratorymeasurements (chemistry, hematology, and urinalysis), 12-leadelectrocardiograms (ECGs), vital signs, and physical examinations.

For TG, TC, HDL-C, LDL-C, calculated non-HDL-C, and VLDL-C, baseline wasdefined as the average of Visit 4 (Week 0) and the preceding lipidqualifying visit (either Visit 3 [Week-1] or if it occurs, Visit 3.1)measurements. Baseline for all other efficacy parameters was the Visit 4(Week 0) measurement.

For TG, TC, HDL-C, LDL-C, calculated non-HDL-C, and VLDL-C, Week 12endpoint was defined as the average of Visit 6 (Week 11) and Visit 7(Week 12) measurements.

Week 12 endpoint for all other efficacy parameters were the Visit 7(Week 12) measurement.

The primary efficacy analysis was performed using a 2-way analysis ofcovariance (ANCOVA) model with treatment as a factor and baseline TGvalue as a covariate. The least-squares mean, standard error, and2-tailed 95% confidence interval for each treatment group and for eachcomparison were estimated. The same 2-way ANCOVA model was used for theanalysis of secondary efficacy variables.

The primary analysis was repeated for the per-protocol population toconfirm the robustness of the results for the intent-to-treatpopulation.

Non-inferiority tests for percent change from baseline in LDL-C wereperformed between >96% E-EPA doses and placebo using a non-inferioritymargin of 6% and a significant level at 0.05.

For the following key secondary efficacy parameters, treatment groupswere compared using Dunnett's test to control the Type 1 error rate: TC,LDL-C, HDL-C, non-HDL-C, VLDL-C, Lp-PLA₂, and apo B. For the remainingsecondary efficacy parameters, Dunnett's test was be used and the ANCOVAoutput were considered descriptive.

The evaluation of safety was based primarily on the frequency of adverseevents, clinical laboratory assessments, vital signs, and 12-lead ECGs.The primary efficacy variable is the percent change in fasting TG levelsfrom baseline to Week 12. A sample size of 194 completed patients pertreatment group provided 90.6% power to detect a difference of 15%between >96% E-EPA and placebo in percent change from baseline infasting TG levels, assuming a standard deviation of 45% in TGmeasurements and a significance level of p<0.05.

Previous data on fasting LDL-C show a difference in percent change frombaseline of 2.2%, with a standard deviation of 15%, between study drugand placebo. A sample size of 194 completed patients per treatment groupprovided 80% power to demonstrate non-inferiority (p<0.05, one-sided) ofthe LDL-C response between >96% E-EPA 4 g daily and placebo, within a 6%margin. To accommodate a 10% drop-out rate from randomization tocompletion of the double-blind treatment period, a total of 648randomized patients was planned (216 patients per treatment group); 702subjects were randomized, as further described below.

Results

Of the 702 randomized subjects, 687 were in the intent-to-treat (“ITT”)population as follows:

-   -   Ultra-pure EPA, 4 g/day: 226 subjects    -   Ultra-pure EPA, 2 g/day: 234 subjects    -   Placebo: 227 subjects

Lipids were extracted from plasma and red blood cell (“RBC”) suspensionsand converted into fatty acid methyl esters for analysis using astandard validated gas chromatography/flame ionization detection method.Fatty acid parameters were compared between EPA treatment groups andplacebo using an ANCOVA model with treatment, gender, type of statintherapy, and presence of diabetes as factors, and the baseline parametervalue as a covariate. LSMs, SEs, and 2-tailed 95% confidence intervalsfor each treatment group and for each comparison were determined.

Baseline characteristics of the three ITT groups were comparable, with61.4% of the ITT subjects being male, 96.3% being white, having a meanage of 61.4 years, a weight of 95.7 kg and a BMI of 32.9 kg/m². ITTsubjects with incomplete fatty acid data at baseline and/or at 12 weekswere excluded from the analyses described below.

Example 3

A phase 1, open-label, crossover, drug-drug interaction study of healthysubjects was conducted to determine the effect—if any—of ethyleicosapentaenoate on the pharmacokinetics (e.g., steady-statepharmacokinetics) of rosiglitazone. The study design allowed forevaluation of potential PK drug interactions between ethyleicosapentaenoate and 2 different drugs metabolized by CYP2C classisozymes, omeprazole (CYP2C19 substrate) and rosiglitazone (CYP2C8substrate), with sequential administration of these drugs separated by4-day washout periods between omeprazole, rosiglitazone, andco-administration of ethyl eicosapentaenoate. Subjects were enrolled toreceive omeprazole on days 1 to 7, rosiglitazone on day 11, ethyleicosapentaenoate on days 12 to 29, omeprazole on days 19 to 25, androsiglitazone on day 29. This report focuses on findings from therosiglitazone portion of the study (days 11 and 29 PK sampling);omeprazole results are reported separately.

Eligibility assessments and clinical laboratory testing were performedwithin a 28-day screening period. All eligible subjects received thesame treatment. Participants received one oral 8-mg tablet ofrosiglitazone 1 hour prior to breakfast on days 11 and 29. On days12-29, subjects received oral doses of 4 g icosapent ethyl (2liquid-filled 1 g gelatin capsules) twice daily, with or following themorning and evening meals. All study drugs were taken with 240 mL water.Rosiglitazone was administered by study personnel at the research unit,and thus compliance calculations were not necessary. icosapent ethyl waseither administered by study personnel during scheduled visits orself-administered by subjects while away from the study site. Complianceto icosapent ethyl (days 12-29) was evaluated by counting andreconciling unused capsules against subject diaries and was calculatedas: (used capsules/total dosing days×4)×100.

Rosiglitazone PK parameters were determined on days 11 and 29 (withoutand with icosapent ethyl, respectively). Blood samples (6 mL) for thedetermination of rosiglitazone plasma concentrations were obtained attime 0 (prior to dose) and at 0.25, 0.5, 0.75, 1, 2, 4, 6, 8, 10, 12,16, and 24 hours after the rosiglitazone dose. Doses selected for studywere based on established PK profiles of both agents. The maximumrecommended dose of rosiglitazone (8 mg) was expected to safely providemaximal exposure, and the 4 g/day dose of icosapent ethyl capsulesrepresents the FDA-approved daily dose.¹ Rosiglitazone may beadministered with or without food. However, to minimize PK variabilityin maximum observed concentration (C_(max)) and time to C_(max)(T_(max)) estimates in the present study, the protocol specified thatrosiglitazone (8 mg) be administered as a single dose 1 hour prior tothe morning meal. The elimination half-life of rosiglitazone is short(3-4 hours, independent of dose), which justifies the 24-hour intervalfor collecting all samples to characterize the PK parameters.

The protocol was approved by an institutional review board (IntegReviewEthics Review Board, Austin, Tex., USA) and was conducted between Feb.3, 2011 and Mar. 21, 2011 at Frontage Clinical Services (a wholly ownedsubsidiary of Frontage Laboratories, Hackensack, N.J., USA). The studycomplied with the ethical principles of Good Clinical Practice and inaccordance with the Declaration of Helsinki. All subjects providedwritten informed consent prior to study entry.

Eligible subjects were healthy nonsmoking men and women between the agesof 19 and 55 years with a body mass index >18 and ≦35 kg/m² and in goodhealth as determined by medical history and medical examination. Womenwho were pregnant, nursing, or planning a pregnancy were excluded;female subjects of childbearing potential were required to use anacceptable method of birth control. Subjects were prohibited fromingesting medications and supplements that contained EPA and/ordocosahexaenoic acid (DHA), fish meals, foods fortified with EPA and/orDHA, lipid-medications and dietary supplements with known or potentiallipid-altering effects including statins, niacin >200 mg/day, fibrates,ezetimibe, and bile acid sequestrants or medications or supplements thatmay influence the measurements of EPA concentrations in plasma untilafter the last PK sample collection. Subjects who required or tookrosiglitazone within 4 weeks prior to the beginning of the study wereexcluded.

Following collection of venous blood samples into pre-chilled glasstubes containing dipotassium ethylenediaminetetraacetic acid (K₂EDTA),plasma was separated by centrifugation for measurement of rosiglitazoneconcentrations using a validated liquid chromatography with tandem massspectrometry (LC-MS/MS) method by Frontage Laboratories, Inc. (Malvern,Pa., USA). Rosiglitazone and rosiglitazone-d₃ were extracted from humanplasma by protein precipitation using acetonitrile and separated byreversed-phase high-performance liquid chromatography (HPLC) with aCadenza CD-C18 column (75×3 mm, 3 μm; Imtakt USA, Philadelphia, Pa.,USA) and Shimadzu HPLC pump and autosampler (Shimadzu, Kyoto, Japan),with a flow rate of 0 5 mL/min at room temperature and an elution timeof 2.8 min. The premixed isocratic mobile phase was acetonitrile:10 mMammonium acetate 45:55 v/v. Rosiglitazone-d₃ was used as the internalstandard and the reference standard was rosiglitazone. Ions weremonitored for rosiglitazone at m/z 358.1-135.0 and for rosiglitazone-d₃at 361.1-138.0 in positive ionization mode using the API4000™ massspectrometer with TurbolonSpray electrospray ion source (AB Sciex,Framingham, Mass., USA) at 450° C. and 4500 V with N₂. The dynamic rangewas 2-800 ng/mL with a lower limit of quantitation of 2 ng/mL. The assayaccuracy (mean determined concentration/nominal concentration) rangedfrom 94.1-108.1% (intra-day) and from 100.9-102.3% (inter-day). Theassay precision (coefficient of variation of the mean determinedconcentration) ranged from 1.0-4.6% (intra-day) and from 4.9-6.2%(inter-day).

PK parameters were derived by noncompartmental analysis using WinNonlinversion 5.0.1 or higher (Pharsight Corporation Inc., Mountain View,Calif., USA) and actual sampling times. The primary PK parametercalculated for rosiglitazone on days 11 and 29 (without and withicosapent ethyl, respectively) was area under the plasmaconcentration-vs-time curve from time zero to infinity (AUC_(0-inf))after a single dose of rosiglitazone, calculated fromAUC_(0-t)+(C_(t)/λ_(z)), where C_(t) was the last observed quantifiableconcentration. Secondary PK end points included C_(max), T_(max), andAUC from time zero to 24 hours (AUC₀₋₂₄). Additional end points includedelimination half-life (t_(1/2)) and apparent terminal elimination rateconstant (K_(el)). Comparisons included only subjects with primary PKparameters available for rosiglitazone from both PK sampling days (PKanalysis population).

A sample size of 30 subjects, with at least 24 subjects completing thestudy, was selected as one that would meet study aims. Theintent-to-treat (ITT) population included all subjects who signed theinformed consent form and were included in the study. The PK populationincluded all subjects who had the primary rosiglitazone PK end pointparameters from days 11 and 29 available. Safety was evaluated for allsubjects who received at least one dose of study drug.

PK parameters were calculated by noncompartmental analysis usingWinNonlin version 5.0.1 (Pharsight Corporation Inc., Mountain View,Calif., USA). For each PK parameter, parametric and/or nonparametricdescriptive statistics were calculated. Parametric statistics includedmean, standard deviation (SD), geometric means, and percent coefficientof variation (% CV). Nonparametric statistics included median and datarange (minimum-maximum). Drug—drug interaction was based on theAUC_(0-inf) of rosiglitazone. Analysis of variance (ANOVA) models wereused for analyzing AUC and C_(max) parameters based on naturallog-transformed values. This included the effects for treatment (withoutor with icosapent ethyl) as a random effect. The estimate of the ratiobetween the two treatments for these parameters and the corresponding90% confidence intervals (CI) for the ratio were obtained byexponentiating the difference in logarithms, and were used to determinewhether a drug—drug interaction of the two treatments (without or withicosapent ethyl) occurred.

Safety evaluations consisted of monitoring adverse events (AEs),clinical laboratory measurements (chemistry, hematology, andurinalysis), vital signs (systolic and diastolic blood pressure, heartrate, respiratory rate and oral body temperature), and physicalexamination findings.

Twenty-eight out of thirty enrolled subjects completed the study. Eachsubject received a single 8-mg oral dose of rosiglitazone, alone andwith 4 g of ethyl eicosapentaenoate in a composition consistent with thepresent disclosure. Mean (standard deviation [SD]) compliance based oncapsule counts for icosapent ethyl for days 12 through 29 was 98.4%(4.2%).

Primary and secondary pharmacokinetic endpoints included area under thecurve versus time (AUC_(0-inf)) and maximum plasma concentration(C_(max)) at steady state for rosiglitazone administered with or withoutthe ethyl eicosapentaenoate (Table 1).

TABLE 1 Statistical Analysis of Drug-Drug Interaction: RosiglitazoneAdministered Alone vs Co-Administration of Rosiglitazone and Ethyl-EPA.Rosiglitazone Rosiglitazone (8 mg) Parameter alone (8 mg) with EPA (4 g)AUC_(0-inf) 3228 (679) ng · h/mL 2921 (677) ng · h/mL AUC₀₋₂₄ 3152 (648)ng · h/mL 2873 (654) ng · h/mL C_(max) 672 (185) ng/mL 673 (170) ng/mLT_(max) 0.8 (0.5, 2.0) h 0.8 (0.5, 2.0) h T_(1/2) 4.4 (0.7) h 4.1 (0.7)h K_(el) 0.16 (0.02) 1/h 0.17 (0.03) 1/h *Mean(SD) displayed for allpharmacokinetic parameters except T_(max), which is displayed as median(min, max). AUC₀₋₂₄ = area under the plasma concentration-vs-time curvefrom time zero to 24 hours; AUC_(0-inf) = area under the plasmaconcentration-vs-time curve from time zero to infinity; C_(max) =maximum observed concentration; K_(el) = apparent terminal eliminationrate constant; T_(1/2) = apparent terminal elimination half-life;T_(max) = time of maximum observed concentration.

A comparison of mean plasma concentrations from 0 hours to 24 hourspost-dose for subjects receiving rosigilitazone with 4 grams/day ofethyl eicosapentaenoate (squares) and without ethyl eicosapentaenoate(diamonds) is shown in FIG. 1.

Least squares geometric mean ratios (at 90% confidence intervals) forAUC_(0-inf) and C_(max) for rosiglitazone with EPA vs. rosiglitazonealone are shown in Table 2 below.

TABLE 2 Statistical Analysis of Drug-Drug Interaction: RosiglitazoneAdministered Alone vs Co-Administration of Rosiglitazone and Ethyl-EPA.LSGM LSGM Ratio LSGM Ratio Parameter Ros Ros + EPA (90% CI) RangeAUC_(0-inf) 3153 2842 0.90 87.00-93.40 C_(max) 647 650 1.01 92.02-109.9LSGM derived from mixed models; LSGM ratios are provided for(Rosiglitazone + EPA)/(Rosiglitazone alone)

Thus, there was no significant difference in either endpoint whenrosiglitazone was administered alone compared to co-administration withethyl eicosapentaenoate.

Concomitant administration of rosiglitazone and ethyl eicosapentaenoatewas safe and well-tolerated. No serious adverse events were reported andno subjects prematurely discontinued the study due to an adverse event.

These data show that, at steady state, ethyl eicosapentaenoate does notinhibit the metabolism of rosiglitazone, and induced no clinicallyrelevant exposure changes when administered concomitantly.

What is claimed is:
 1. A method of reducing triglycerides in a subjecton rosiglitazone therapy, the method comprising administering to thesubject a pharmaceutical composition comprising at least about 80%, byweight of all fatty acids (and/or derivatives thereof) present, ethyleicosapentaenoate.
 2. The method of claim 1, wherein the subject has afasting baseline triglyceride level of about 200 mg/dl to 499 mg/dl. 3.The method of claim 1, wherein the subject has a fasting baselinetriglyceride level of at least 500 mg/dl.
 4. The method of claim 1,wherein triglycerides are reduced in the subject with no increase in anLDL-C level in the subject.
 5. The method of claim 4, wherein thereduction in triglycerides and the no increase in LDL-C level is incomparison to baseline or to a second subject or subject group that hasreceived rosiglitazone but not the ethyl eicosapentaenoate.
 6. Themethod of claim 1, wherein ethyl eicosapentaenoate represents at leastabout 90%, by weight of all fatty acids (and/or derivatives thereof)present.
 7. The method of claim 6, wherein ethyl eicosapentaenoaterepresents at least about 95%, by weight of all fatty acids (and/orderivatives thereof) present.
 8. The method of claim 7, wherein ethyleicosapentaenoate represents at least about 96%, by weight of all fattyacids (and/or derivatives thereof) present.
 9. The method of claim 1,wherein docosahexaenoic acid and its esters represent no more than about20%, by weight of all fatty acids (and/or derivatives thereof) presentin the pharmaceutical composition or capsule.
 10. The method of claim 9,wherein docosahexaenoic acid and its esters represent no more than about10%, by weight of all fatty acids (and/or derivatives thereof) presentin the pharmaceutical composition or capsule.
 11. The method of claim10, wherein docosahexaenoic acid and its esters represent no more thanabout 5%, by weight of all fatty acids (and/or derivatives thereof)present in the pharmaceutical composition or capsule.
 12. The method ofclaim 11, wherein docosahexaenoic acid and its esters represent no morethan about 3%, by weight of all fatty acids (and/or derivatives thereof)present in the pharmaceutical composition or capsule.
 13. The method ofclaim 1, wherein the rosiglitazone therapy comprises administering tothe subject about 4 mg or about 8 mg of rosiglitazone per day.
 14. Amethod of reducing triglycerides in a subject on rosiglitazone therapy,the method comprising administering to the subject about 4 g per day ofethyl eicosapentaenoate.
 15. The method of claim 14, wherein the subjecthas a fasting baseline triglyceride level of about 200 mg/dl to 499mg/dl.
 16. The method of claim 14, wherein the subject has a fastingbaseline triglyceride level of at least 500 mg/dl.
 17. The method ofclaim 14, wherein triglycerides are reduced in the subject with noincrease in an LDL-C level in the subject.
 18. The method of claim 17,wherein the reduction in triglycerides and the no increase in LDL-Clevel is in comparison to baseline or to a second subject or subjectgroup that has received rosiglitazone but not the ethyleicosapentaenoate.
 19. The method of claim 14, wherein ethyleicosapentaenoate comprises at least about 80%, by weight of all fattyacids (and/or derivatives thereof) present.
 20. The method of claim 19,wherein ethyl eicosapentaenoate represents at least about 90%, by weightof all fatty acids (and/or derivatives thereof) present.
 21. The methodof claim 20, wherein ethyl eicosapentaenoate represents at least about95%, by weight of all fatty acids (and/or derivatives thereof) present.22. The method of claim 21, wherein ethyl eicosapentaenoate representsat least about 96%, by weight of all fatty acids (and/or derivativesthereof) present.
 23. The method of claim 14, wherein docosahexaenoicacid and its esters represent no more than about 20%, by weight of allfatty acids (and/or derivatives thereof) present in the pharmaceuticalcomposition or capsule.
 24. The method of claim 23, whereindocosahexaenoic acid and its esters represent no more than about 10%, byweight of all fatty acids (and/or derivatives thereof) present in thepharmaceutical composition or capsule.
 25. The method of claim 24,wherein docosahexaenoic acid and its esters represent no more than about5%, by weight of all fatty acids (and/or derivatives thereof) present inthe pharmaceutical composition or capsule.
 26. The method of claim 25,wherein docosahexaenoic acid and its esters represent no more than about3%, by weight of all fatty acids (and/or derivatives thereof) present inthe pharmaceutical composition or capsule.
 27. The method of claim 14,wherein the rosiglitazone therapy comprises administering to the subjectabout 4 mg or about 8 mg of rosiglitazone per day.
 28. A method ofreducing a risk of a cardiovascular event in a subject on rosiglitazonetherapy, the method comprising administering to the subject apharmaceutical composition comprising at least about 80%, by weight ofall fatty acids (and/or derivatives thereof) present, ethyleicosapentaenoate.
 29. The method of claim 28, wherein the rosiglitazonetherapy comprises administering to the subject about 4 mg or about 8 mgof rosiglitazone per day.
 30. A method of reducing a risk of acardiovascular event in a subject on rosiglitazone therapy, the methodcomprising in a subject on rosiglitazone therapy, the method comprisingadministering to the subject about 4 g per day of ethyleicosapentaenoate.
 31. The method of claim 30, wherein the rosiglitazonetherapy comprises administering to the subject about 4 mg or about 8 mgof rosiglitazone per day.